Comparison of ZyFISH and progeny testing assay for zygosity determination in C57BL/6-tga20 littermates.

Six C57BL/6-tga20 transgene-positive mice with predicted zygosity (Figure 2A) according to zyFISH (ZyFISH Result) were each crossed with a C57BL/6 non-transgenic mate. The resulting offspring were genotyped via conventional PCR (tg+ = number of transgene-positive offspring, n = total number of offspring). All mice predicted by zyFISH to be tg/tg had 100% transmittance of the transgene to their offspring, whereas predicted hemizygous mice transmitted the transgene with at a rate of ∼50% (PCR positive F1 offspring). For each of the tg/tg mice we calculated the probability of a tg/0 mouse to sire this many tg+ offspring in succession, by 0.5n (P/(tg/0)). Not applicable (N/A).